How do I establish a strain after the transgenic Founder mice are obtained?

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Model Description

After the Founder mice are obtained, a transgenic strain can be established according to the following procedure. <img src="default/20180401/441314f44830acd8b960fab7a553a91b.jpg" title="FAQ-CS5-01-xg.jpg" alt="FAQ-CS5-01-xg.jpg" class="img-responsive"> <p style="text-align: justify;">Figure 1. Procedure to establish a mouse strain with a randomly inserted transgene. <table><tbody><tr class="firstRow"><td valign="top" style="word-break: break-all;"><ul class=" list-paddingleft-2"> <li><p style="text-align: justify;">Each Founder mouse is respectively mated with a wild-type mouse (such as C57BL/6J)</li> <li><p style="text-align: justify;">The mating among Founder mice is prohibited</li> <li><p style="text-align: justify;">Without a well-defined method for the identification of homozygous and heterozygous mice, the mating among heterozygous mice is prohibited. </li> <li><p style="text-align: justify;">It is recommended to mate the positive mice in each generation with wild-type mice, and it is not recommended to use the wild-type mice from the same litter</li> <li><p style="text-align: justify;">It is recommended to mate each positive mouse in the F1 generation with wild-type mice until the F2 generation or above is obtained. The experiment can be initialized after the expression of transgene is identified.</li> </ul></td></tr></tbody></table> <br> <p style="text-align: justify;"><strong>Notes about transgenes mediated by the Piggybac transposase system: </strong> <p style="text-align: justify;">The Piggybac transposase tends to insert the target fragment into a transcriptionally active region, thus greatly increasing the probability of obtaining Founder mice with positive expression of the target gene. However, the insertion of transgenes mediated by the Piggybac transposase system may cause gene integration at multiple sites.Due to the existence of integration at multiple sites, the following phenomena will occur during the subsequent breeding of Founder mice along with the separation of multiple integration sites: 1. The proportion of mice with positive results of genotype identification is higher than that of single site insertion; 2. In progeny mice with positive expression of the target gene, the level of expression is lower than that in the Founder mice (the amount of reduction is dependent on the number and location of integration sites in Founder mice) and is inconsistent among different progeny mice.Therefore, it is recommended that each Founder mouse be backcrossed with wild-type mice for at least 2–3 generations to obtain transgenic mice with single site integration and stable expression of the exogenous gene. <br>
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